Total lipids from stationary phase promastigotes (1.0 x 108 cells/sample) were extracted using the Bligh & Dyer method [65 (link)]. Commercial non-indigenous lipid standards were added to cell lysates as internal standards at the time of lipid extraction (1.0 x 108 molecules/cell for 14:0/14:0-PE, 5.0 x107 molecules/cell for 14:0/14:0-PC, and 1.0 x 108 molecules/cell for 16:0/16:0-PI). These lipid standards are absent from L. major promastigotes. Determination of lipid families by electrospray ionization mass spectrometry (ESI-MS) was carried out by a Thermo Vantage TSQ instrument applying precursor ion scan of m/z 196 for PE, precursor ion scan of m/z 241 for PI and IPC in the negative-ion mode, and precursor ion scan of m/z 184 for PC in the positive-ion mode [66 (link)]. Individual lipid species and their structures were also confirmed by high resolution mass spectrometry performed on a Thermo LTQ Obitrap Velos with a resolution of 100,000 (at m/z 400). All lipidomic analyses were performed five times.
Ltq obitrap velos
Manufactured by Thermo Fisher Scientific
The LTQ Orbitrap Velos is a high-performance hybrid mass spectrometer that combines a linear ion trap (LTQ) and an Orbitrap mass analyzer. It provides high mass resolution, mass accuracy, and sensitivity for a wide range of applications in proteomics, metabolomics, and small molecule analysis.
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Lab products found in correlation
2 protocols using ltq obitrap velos
1
Lipidomic Analysis of Leishmania major
2
Lipid Extraction and Profiling of Leishmania Promastigotes
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Total lipids from stationary phase promastigotes (1.0 x 108 cells/sample) were extracted using the Bligh & Dyer method [57 (link)]. Commercial non-indigenous lipid standards were added to cell lysates as internal standards at the time of lipid extraction (1.0 x 108 molecules/cell for 14:0/14:0-PE, 5.0 x107 molecules/cell for 14:0/14:0-PC, and 1.0 x 108 molecules/cell for 16:0/16:0-PI). These lipid standards are absent from L. major promastigotes. Determination of lipid families by electrospray ionization mass spectrometry (ESI-MS) was carried out by a Thermo Vantage TSQ instrument applying precursor ion scan of m/z 196 for PE, precursor ion scan of m/z 241 for PI and IPC in the negative-ion mode, and precursor ion scan of m/z 184 for PC in the positive-ion mode [58 (link)]. Individual lipid species and their structures were also confirmed by high resolution mass spectrometry performed on a Thermo LTQ Obitrap Velos with a resolution of 100,000 (at m/z 400). All lipidomic analyses were performed five times.
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